LOCUS       sequence-379396-        8812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible vector with BsaI cloning site and Cre-2A-Cas9 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8812)
  AUTHORS   Merle N, Elmshauser S, Strassheimer F, Wanzel M, Konig AM, Funk J, 
            Neumann M, Kochhan K, Helmprobst F, Pagenstecher A, Nist A, 
            Mernberger M, Schneider A, Braun T, Borggrefe T, Savai R, Timofeev 
            O, Stiewe T
  TITLE     Monitoring autochthonous lung tumors induced by somatic CRISPR gene 
            editing in mice using a secreted luciferase.
  JOURNAL   Mol Cancer. 2022 Oct 3;21(1):191. doi: 10.1186/s12943-022-01661-2.
  PUBMED    36192757
REFERENCE   2  (bases 1 to 8812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Cancer."; date: "2022-10-3"; volume: "21(1):191. doi"; pages: " 
            10.1186/s12943-022-01661-2"
COMMENT     Sequence Label: pShuttle.CC9
FEATURES             Location/Qualifiers
     source          1..8812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        713..1016
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1017..1220
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1171..1191
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1217..1241
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1309..1329
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1330..2355
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             2365..2418
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2422..2487
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2494..2514
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2539..6639
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2539..6639
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6640..6687
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6715..6732)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6721..6928
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(6937..7036)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7053..7072)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7054..7072)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7077..7093)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7077..7093)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7195..7214)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7206..8015
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7273..7292)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      8162..8750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8651..8670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcacc atggaggcta gagacctaga gcgatcgctc gcggtctcac cgcgttacat
      721 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      781 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      841 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      901 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      961 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1021 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1081 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1141 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     1201 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
     1261 gctgttttga cctccataga agacaccgac tctagaggat ctaccatgcc caagaagaag
     1321 aggaaggtgt ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca
     1381 acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct
     1441 gagcatacct ggaaaatgct tcagtccgtt tgccggtcgt gggcggcatg gtgcaagttg
     1501 aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt
     1561 caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt
     1621 catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg
     1681 cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa
     1741 cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata
     1801 cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc
     1861 aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc
     1921 agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact
     1981 aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg
     2041 ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact
     2101 cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac
     2161 tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat
     2221 atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta
     2281 aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg
     2341 ctggaagatg gcgatggaag cggagagggc agaggaagtc tgctaacatg cggtgacgtc
     2401 gaggagaatc ctggacctat ggactataag gaccacgacg gagactacaa ggatcatgat
     2461 attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc
     2521 cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct
     2581 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     2641 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     2701 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     2761 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     2821 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     2881 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     2941 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     3001 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     3061 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     3121 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     3181 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     3241 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     3301 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     3361 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     3421 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     3481 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     3541 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     3601 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     3661 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     3721 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     3781 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     3841 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     3901 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     3961 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     4021 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     4081 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     4141 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     4201 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     4261 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     4321 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     4381 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     4441 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     4501 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     4561 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     4621 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     4681 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     4741 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     4801 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     4861 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     4921 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     4981 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     5041 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     5101 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     5161 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     5221 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     5281 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     5341 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     5401 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     5461 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     5521 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     5581 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     5641 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     5701 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     5761 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     5821 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     5881 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     5941 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     6001 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     6061 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     6121 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     6181 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     6241 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     6301 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     6361 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     6421 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     6481 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     6541 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     6601 tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa
     6661 aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag agctcgctga tcagcctcga
     6721 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6781 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6841 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6901 gggaagagaa tagcaggcat gctggggaat tctagaaccc agctttcttg tacaaagttg
     6961 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     7021 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     7081 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     7141 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     7201 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     7261 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     7321 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     7381 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     7441 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     7501 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     7561 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     7621 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     7681 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     7741 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     7801 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     7861 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     7921 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     7981 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     8041 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     8101 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8161 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8221 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8281 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     8341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8701 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8761 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
